(A) SOP6 and gliotoxin induce cell detachment. Shown are microscopic
images of HCT116 cells, taken 3 hours after treatment with 10 μM of the
indicated ETPs. Note that when the cells detach, as in the top two panels, they
appear more refractile and have a tendency to clump. (B) c-Myc mRNA expression,
quantified by qRT-PCR, in HCT116 cells harvested 4 hours post treatment. 1
μM actinomycin D was included as positive control, and 10 μM other
compounds were used. Data are normalized to GAPDH. Error bars mean ± s.d.
from three independent experiments measured in triplicate each. * P<0.05,
** P<0.01 (C) HCT116 cells were treated for 6 hours with indicated
compounds, and cell lysates were fractionated by SDS–PAGE and
immunoblotted with antibodies against NFE2L1, NFE2L2, Cul1 and GAPDH (loading
control for C and D). (D) Same as C, except that antibodies against XBP1, CHOP,
PERK and Bip were used. See also Figure S2, Figure S3 and Figure S4.